HPLC-MS/MS同时检测大鼠体内三聚氰胺和三聚氰酸含量  

Simultaneous determination of melamine and cyanuric acid in rat by HPLC-MS/MS

作　　者：何君[1] 高虹[1] 李岩[1] 秦超[1] 陈威[1] 舒健明[1] 孙继超[1] 王皎[1] 秦川[1] 

机构地区：[1]中国医学科学院医学实验动物研究所北京协和医学院比较医学中心,北京100021

出　　处：《中国食品卫生杂志》2012年第4期322-329,共8页Chinese Journal of Food Hygiene

基　　金：卫生部公益性行业科研专项经费(200902009-04)

摘　　要：目的建立HPLC-MS/MS同时检测大鼠血浆和组织中三聚氰胺和三聚氰酸含量的方法。方法以同位素标记的三聚氰胺和三聚氰酸作为内标物,大鼠血浆和组织经二氯甲烷-乙腈前处理后,经色谱柱分离,串联离子阱质谱采用ESI源按运行时间段切换正、负离子模式进行测定。结果在0.3～75μg/ml范围内,三聚氰酸和三聚氰胺具有良好的线性(r≥0.998)。在0.9、5.0、50μg/ml的给药大鼠的血浆样本中,三聚氰酸和三聚氰胺的回收率分别为97.4%～99.4%、96.8%～98.9%,两者的变异系数为2.43%～6.35%(日内)、1.70%～4.12%(日间)和0.30%～9.17%(日内)、0.75%～3.39%(日间);在0.9、5.0、50μg/ml的给药大鼠肝组织样本中,三聚氰酸和三聚氰胺的回收率分别为90.8%～104.6%、91.7%～106.0%,对应的变异系数为3.70%～5.14%(日内)、2.84%～4.50%(日间)和1.31%～2.21%(日内)、1.12%～3.36%(日间)。结论该方法样品前处理简单,基质干扰小,分析效率高,具有良好的精密度、准确度和特异性,是进行三聚氰酸、三聚氰胺在动物体内的组织代谢和毒性研究的理想分析方法。Objective To establish a method for simultaneous determination of melamine and cyanuric acid in rat by HPLC-MS/MS. Methods Using isotope-labelled melamine and cyanuric acid as internal standard, rat plasma and tissue samples were pretreated with dichloromethane-acetonitrile, and separated by chromatographic column switching technique. Detection was carried out by HPLC coupled with electrospray ionization （ESI） working on segment ESI- and ESI＋ switching mode. Results The calibration curve of melamine and cyanuric acid were linear （ r ≤0. 998） in the range of 0.3-75 μg/ml. The average recovery rates of melamine and eyanuric acid （ added with 0.9, 5.0 and 50 μg/ml） in rat plasma were 97.4%-99.4% and 96. 8%-98.9%, respectively; while the relative standard deviations （RSD） of the method were 2.43% -6.35% （inter-day） , 1.70% -4. 12% （intra-day） , 0. 30% -9.17% （inter-day） and 0. 75% -3.39% （intra-day） , respectively. The average recovery rates of rat liver tissue （added with 0. 9, 5.0 and 50 μg/ml） were 90. 8%-104.6% and 91.7%-106.0%, respectively; while the relative standard deviations （RSD） of the method were 3.70%-5.14% （inter-day）, 2.84%-4.50% （intra-day）, 1.31%-2.21% （inter-day） and 1.12%-3.-36% （intraday）, respectively. Conclusion The method was simple in sample pretreatmcnt, less matrix interference, efficient, precise, accurate and specific. It was ideal to study the mechanism of metabolic behavior and toxicology of melamine and cyanuric acid in animals.

关 键 词：三聚氰酸 三聚氰胺 血浆 组织 HPLC-MS/MS 

分 类 号：R114[医药卫生—卫生毒理学]